Adhesive sealant composition

ABSTRACT

This invention is related to an adhesive composition which may be used to bond or seal tissue in vivo. The adhesive composition is readily formed from a two component mixture which includes a first part of a protein, preferably a serum albumin protein, in an aqueous buffer having a pH in the range of about 8.0-11.0 and a second part of a water-compatible or water-soluble bifunctional crosslinking agent. When the two parts of the mixture are combined, the mixture is initially a liquid which cures in vivo on the surface of tissue in less than about one minute to give a strong, flexible, pliant substantive composition which bonds to the tissue and is absorbed in about four to sixty days. The adhesive composition may be used either to bond tissue, to seal tissue or to prevent tissue adhesions caused by surgery.

Notice: More than one reissue application has been filed for the reissueof U.S. Pat. No. 5,583,114. The reissue applications are applicationSer. No. 10/293,989, filed Nov. 14, 2002, and application Ser. No.09/185,732, filed Nov. 4, 1998, now U.S. Pat. Re 38,158, all of whichare reissues of U.S. Pat. No. 5,583,114.

This application is a continuation of reissue application Ser. No.09/185,732, filed Nov. 4, 1998, now U.S. Pat. No. RE 38,158, issued Jun.24, 2003, which is a reissue of U.S. Pat. No. 5,583,114.

The present invention is generally related to an adhesive sealantcomposition which may be used to bond or seal tissue in vivo and isparticularly related to a two component, liquid adhesive compositionwhich is mixed together as it is applied to tissue and then cured invivo in order to bond tissue, to seal tissue to prevent or controlpulmonary system air leaks, or to prevent tissue adhesions caused bysurgery.

BACKGROUND

A variety of techniques have been used to bond or seal tissue. Forexample, different types of tissues have been mechanically bound orsealed with a number of procedures, materials and methods includingsutures, staples, tapes and bandages. In some applications, thesematerials are made of absorbable materials which are intended to bondand/or seal tissue as it heals and then to be absorbed over a period oftime.

The common use of a medical adhesive to “tissue glue” has not foundwidespread application. To date, some adhesive materials are known whichmay be used to adhere or stick tissue such as skin. For example,cyanoacrylate adhesives such as HISTOACRYL adhesive available from B.Braun, Melsungen, Germany or VETBOND tissue adhesive available from 3M,St. Paul, Minn. may be used to bond tissue. In addition to cyanoacrylateadhesives, other types of materials have been reported to adhere orstick to skin. For example, U.S. Pat. No. 4,839,345 to Doi et al.reports a hydrated crosslinked protein adhesive gel that is used as acataplasm or cosmetic mask that will externally adhere to skin but canbe easily removed or pulled off and then readhered to the skin. Othercrosslinked protein hydrogels have been reported to serve as aproteinaceous substrate to deliver therapeutic agents such as enzymes ordrugs through skin or mucous membranes. See, for example, InternationalPatent Application Ser. No. PCT/US93/07314 filed Aug. 4, 1993. Stillother materials have been used as hemostatic agents to stop or preventbleeding. In particular, mixtures of fibrinogen and thrombin such asTISSEEL sealant available from Immuno AG, Vienna, Austria or BERIPLAST-Phemostatic agent or sealant available from Behringwerke, Marburg,Germany, have been used in vascular surgery to seal tissue such as bloodvessels and thus prevent blood leakage.

In sum, there are few available adhesive compositions that havesufficient strength, biocompatibility and bioabsorbability as well asother desired properties that would allow such compositions to bereadily used in current medical procedures or practices. Theunavailability of a suitable tissue adhesive or sealant may be relatedto the stringent requirements that a suitable, useful tissue adhesivemust meet. Importantly, a tissue adhesive must provide substantialbonding strength for either internal or external tissues. The adhesiveshould be made of a biocompatible material which does not interfere withnormal healing or regeneration processes. A suitable tissue adhesivemust also be easily administered in a liquid form and then rapidlycured, ideally in less than a minute, one applied. In addition, a tissueadhesive must remain flexible, pliant and have good mechanical strengthafter being cured. Finally, a tissue adhesive must be completelyabsorbed or broken down in vivo, without producing an allergic response,adverse tissue reaction or systemic toxic effects, in an acceptable timeperiod. Preferably a suitable adhesive would also be readily absorbedafter it is applied.

SUMMARY OF THE INVENTION

The present invention is a nontoxic, absorbable adhesive sealantcomposition which may be used to bond and/or seal tissue. The adhesivecomposition is readily formed from a two component mixture whichincludes a first part of a protein, preferably a serum protein such asalbumin, in an aqueous buffer having a pH in the range of about 8.0-11.0and a second part of a water-compatible or water-soluble bifunctionalcrosslinking agent. When the two parts of the mixture are combined, themixture is initially liquid. The combined mixture then cures in vivo onthe surface of tissue in less than about one minute to give a strong,flexible, pliant substantive composition which securely bonds to thetissue and is readily absorbed in about four to sixty days, preferablyin about four to twenty-eight days.

In a preferred embodiment of the invention, an adhesive sealantcomposition is formed from a two part mixture that includes a proportionof a volume of a buffered basic serum albumin protein solution to avolume of a polyethylene glycol disuccinimidoyl succinate crosslinkingagent in a range of from about 1:10 parts albumin solution by volume toabout 10:1 parts by volume crosslinking agent. In order to facilitatethe mixing of the two parts of the present adhesive composition, thevolume to volume ratio of albumin solution to crosslinking agent ispreferably a ratio of 1:1.

Preferred serum albumin proteins are selected to prevent adverse tissueor unwanted immunological responses. When the present adhesive mixtureis used to bond or seal human tissue, a preferred serum albumin ispurified human serum albumin which has been sterilized, dialyzed with abasic buffer having a pH value of about 8.0-11.0 concentrated byultrafiltration through a membrane having about a 50,000 molecularweight cut-off to yield a concentrated, buffered aqueous mixture havingabout 20-60 wt/vol %, preferably about 35-45 wt/vol %, human serumalbumin.

Preferred bifunctional crosslinking agents include polyethylene glycolderived crosslinking agents having a molecular weight (weight average)in a range of about 1,000-15,000 and preferably in a range of about2,006-4,000. When the molecular weight of the crosslinking agent is inthe range of about 1,000-5,000 the crosslinking agent is generallydissolved in water at a concentration of about 50-300 mg/ml. Similarly,when the molecular weight of the crosslinking agent is in the range ofabout 5,000-15,000 the crosslinking agent is generally dissolved inwater at a concentration in the range of about 300-800 mg/ml.

The adhesive composition of this invention may be used in a variety ofapplications. Some applications include using the adhesive sealantcomposition to bind tissue together either as an adjunct to or as areplacement of sutures, staples, tapes and/or bandages. In anotherapplication, the present adhesive may be used to prevent post-surgicaladhesions. In this application, the adhesive composition is applied andcured as a layer on surfaces of internal organs or tissues in order toprevent the formation of adhesions at a surgical site as the site heals.Additional applications include sealing tissues to prevent or controlblood or other fluid leaks at suture or staple lines as well as toprevent or control air leaks in the pulmonary system.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graphical representation of a measured peel force of anadhesive composition of this invention.

FIG. 2 is a graphical representation of peel force measurements ofdifferent adhesive composition samples which are used to adhere excisedguinea pig skin strips together.

FIG. 3 is a schematic diagram of an apparatus used to measure burststrength of an adhesive sealant composition.

DETAILED DESCRIPTION

The present invention is related to an adhesive composition which hashigh mechanical strength, flexibility, fast cure rate and sufficientadhesion needed to bond and/or seal tissue in vivo. The adhesivecomposition is made of two components, a buffered basic protein solutionand a bifunctional crosslinking agent. The buffered protein solution andthe bifunctional crosslinking agent are typically prepared usingcommercially available materials and established synthetic methods. Theuse of known, commercially available materials in the preparation of theadhesive composition provides a benefit in the practice of thisinvention because most of these materials generally have a history ofclinical safety and/or use.

Suitable proteins for use in the present adhesive composition includenonimmunogenic, water soluble proteins. Serum lipoproteins areparticularly well suited for this purpose because these proteins bind tolipids and also exhibit a relatively high elasticity in the natured orsemi-natured state. These properties are believed to provide a curedmatrix which is strong as well as pliant and elastic. Other solubleproteins, in addition to serum lipoproteins, are also suitable for usein the present invention. Aqueous mixtures of proteins such asderivatives of elastin, fibrinogen and collagen may be used in thepresent invention.

Preferred buffered protein solutions which may be used in the presentadhesive composition include concentrated aqueous serum albumin proteinmixtures that are buffered to a pH of between about 8.0-11.0 where thebuffer concentration is in a range of about 0.01-0.25 molar. Suitablebuffer systems include buffers which are physiologically and/orclinically acceptable such as known carbonate or phosphate buffersystems, provided the buffer does not adversely react with or otherwisealter the crosslinking agent. A preferred buffer system is acarbonate/bicarbonate buffer system at a pH value of about 9.0-10.5 at aconcentration in the range of 0.05-0.15 molar.

Serum albumin protein is readily isolated from serum using knownisolation processes. In addition, it is possible to produce human serumalbumin from genetically transformed cells. See, for example, thereports of Quirk et al., Biotechnology and Applied Biochemistry,11:273-287 (1989), Kalman et al., Nucleic Acids Research, 18:6075-6081(1990), Sleep et al., Biotechnology, 8:42-46 (1990), and Sijmons et al.,Biotechnology, 8:217-221 (1990). The ability to produce human serumalbumin recombinantly provides the benefit that protein produced by thismethod will be free of pathogens, viruses or other contaminants thatmight contaminate albumin that is isolated directly from serum.

When used in the present buffered mixtures it has been found that theserum albumin is not denatured. Because the albumin is not denaturedbefore it is used it is believed that the albumin proteins retain theirnatured, coiled conformation and thus, after being crosslinked duringthe curing process to provide a gel-like solid, the cured adhesiveretains sufficient flexibility to provide a suitable adhesive matrix.

A variety of suitable crosslinking agents may be used in the presentinvention. Preferred crosslinking agents include a polyethylene glycolor polyoxyethylene chain portion (—PEG—), an activated leaving groupportion (—G) and a linking moiety (—LM—) which binds the —PEG— portionand the leaving group portion —G. Crosslinking agents include compoundsof the formulaG—LM—PEG—LM—Gin which —PEG— is a diradical fragment represented by the formula—O—(CH₂—CH₂—O—)_(a)—where a is an integer from 20-300; —LM— is also a diradical fragmentsuch as a carbonate diradical represented by the formula, —C(O)—, amonoester diradical represented by the formula, —(CH₂)_(b)C(O)— where bis an integer from 1-5, a diester diradical represented by the formula,−C(O)—(CH₂)_(c)—C(O)— where c is an integer from 2-10 and where thealiphatic portion of the radical may be saturated or unsaturated, adicarbonate represented by the formula —C(O)—O—(CH₂)_(d)—O—C(O)— where dis an integer from 2-10, or an oligomeric diradical represented by theformulas —R—C(O)—, —R—C(O)—(CH₂)_(c)—C(O)—, or—R—C(O)—O—(CH₂)_(d)—O—C(O)— where c is an integer from 2-10, d is aninteger from 2-10, and R is a polymer or copolymer having 1-10 monomericlactide, glycolide, trimethylene carbonate, caprolactone and p-dioxanonefragments; and —G is a leaving group such as a succinimidyl, maleimidyl,phthalimidyl, or alternatively, nitrophenyl, imidazolyl or tresylleaving groups.

The —PEG— portion of the crosslinking agent is preferably derived fromcommercially available compounds having a weight average molecularweight in the range of about 1,000-15,000, preferably having a weightaverage molecular weight in the range of about 2,000-4,000. Thesecompounds have been used in different types of biomedical materialsbecause they have been demonstrated to be non-toxic as well as rapidlyexcreted from the body when the molecular weight is below about 30,000.

The leaving group, —G, portion of the crosslinking agent is an activatedleaving group which allows the crosslinking agent to react or chemicallybind to free primary or secondary amine groups of a protein. Suitableleaving groups include succinimidyl, other imides such as maleimidyl andphthalimidyl, heterocyclic leaving groups such as imidazolyl, aromaticleaving groups such as a nitrophenyl, or fluorinated alkylsulfoneleaving groups such as tresyl (CF₃—CH₂—SO₂—O—). A preferred leavinggroup is the succinimidyl group because studies of the mutagenicity,oncogenicity and teratogenicity of this group suggest that the smallamount of this activating group which is released as the crosslinkingreaction and/or the adhesive composition cures does not present a localor systemic toxicology risk.

When used in the present composition the linking moiety, —LM—, may beseveral different types of divalent compounds. For example, commerciallyavailable compounds having the —PEG— portion and the —G portion linkedwith a saturated dicarboxylic acid such as succinic acid to give asaturated diester linking moiety. Alternatively, an unsaturateddicarboxylic acid such as fumaric, maleic, phthalic or terephthalic acidmay be used to give an unsaturated diester linking moiety.Alternatively, the linking moiety may be a readily hydrolyzablecompounds such as oligomer derivatives of polylactic acid, polyglycolicacid, polydioxanone, polytrimethylene carbonate, or polycaprolactone aswell as copolymers made using suitable monomers of these listedpolymers.

In another embodiment of this invention an activated leaving group maybe attached directly to a carbonate ester of polyethylene glycol. Inthis embodiment the linking moiety, —LM—, would be a carbonate group,—C(O)— between the —PEG— and —G portions of the crosslinking agent. Instill other embodiments of this invention the linking moiety may be adicarbonate such as ethylene carbonate which is prepared by linking the—PEG and —G portions with ethylene bischloroformate.

The crosslinking agents may be prepared using known processes,procedures or synthetic methods such as the procedures reported in U.S.Pat. Nos. 4,101,380 or 4,839,345, the procedure reported inInternational Application Ser. No. PCT/US90/02133 filed Apr. 19, 1990 orthe procedure reported by Abuchowski et al., Cancer Biochem. Biophys.,7:175-186 (1984). Briefly, polyethylene glycol and a suitable acidanhydride are dissolved in a suitable polar organic solvent in thepresence of base and refluxed for a period of time sufficient to form apolyethylene glycol diester diacid. The diester diacid is then reactedwith a leaving group such as an N-hydroxy imide compound in a suitablepolar organic solvent in the presence of dicyclohexylcarbodiimide orother condensing agents and stirred at room temperature to form thedesired bifunctional crosslinking agent.

Alternatively, polyethylene glycol and a suitable dicarboxylic acidchloride or bischloroformate may be dissolved in a suitable polarorganic solvent for a period of time sufficient to form the mixed acidchloride polyethylene glycol ester or mixed chloroformate polyethyleneglycol ester. The mixed esters may then be reacted with a compound suchas an N-hydroxy imide compound in a suitable polar organic solvent andstirred at an elevated temperature for a period of time sufficient toform the desired bifunctional crosslinking agent.

It has also been found that the cure time of the present adhesivecompositions may be tailored by use of buffers having different pHvalues. For example, by varying the pH of the buffer it is possible tochange the cure rate time from about 10 seconds to less than about 10minutes. Briefly, mixing concentrated aqueous serum albumin andcrosslinking agent mixtures with higher concentrations of bufferprovides the fastest cure times. It has also been found that higherconcentrations of protein and crosslinking agent provide a relativelystronger, cured matrix. However, if the mixtures are too concentratedand viscosity becomes too great, these adhesive compositions are not asreadily applied or may provide adhesives with undesired properties. Forexample, mixtures which are too viscous may not be readily applied usingavailable applicators such as syringes or spray apparatus. In addition,if the concentration of crosslinking agent is too high, the resultingcured adhesive matrix may swell to such an extent that the strength ofthe matrix in the presence of water or other fluids is lowered. Further,ability to adequately mix the two components using injecting and/orspraying apparatus may be reduced.

The two component adhesive composition of the present invention may beapplied to tissue in a number of different ways. For example, theadhesive may be quickly mixed together and then applied using commonapplicators. Alternatively the two components may be mixed together andthen applied as spray. In another application method, the two parts ofthe adhesive are added to a dual syringe. The two barrels of the syringeare attached to a “Y” connect which is fitted to a spiral mixer nozzle.As the two components are pressed out of the syringe, they are mixed inthe nozzle and may be directly applied to the tissue as needed in arelatively uniform, controlled manner. Alternatively, a spray nozzletip, such as a TISSEEL spray tip sold by Immuno AG, Vienna, Austria foruse with a two-component fibrin sealant kit, may be used in place of thespiral mixer nozzle. In this application, a fine spray of the adhesivecomposition is deposited on tissue as the plungers of the syringe aredepressed.

The adhesive composition of the present invention may be used in avariety of current medical procedures an practices. In one application,the present adhesive composition may be used to eliminate orsubstantially reduce the number of sutures normally required usingcurrent practices as well as eliminate the need for subsequent removalof certain sutures. In another application, this adhesive compositionmay be used to attach skin grafts and to position tissue flaps or freeflaps during reconstructive surgery. In still another application, thisadhesive composition may be used to close gingival flaps in periodontalsurgery. In all of these applications, the present adhesive compositionis a thin layer of cured material which is effectively sandwichedbetween two adjacent layers of living tissues. Due to bioabsorbabilityand lack of toxicity of the adhesive composition, the healing andsubsequent reattachment of the two layers of tissue to each other is nothampered.

In addition to the use of the present adhesive composition as anadhesive per se, the present composition may also be used as a sealant.When used in this application, this composition may be used to preventair leaks now associated with pulmonary surgery or to inhibit or preventbleeding in other surgical procedures. When used in this manner, theunderlying tissue may be coated with a relatively thick layer ofadhesive since the tissue itself needs to only heal on one side. Theother side of the of the adhesive, when cured, simply presents alubricous gel which will be absorbed in vivo in a relatively shortperiod of time from about four to sixty days. In view of this propertyof the present adhesive composition, it may also be used to preventunwanted tissues adhesions which are associated with current surgicalprocedures.

EXAMPLES

The following examples are intended to describe and illustrate thepractice of the claimed invention. The examples, however, should not beconstrued to limit the scope of the present invention which is definedby the appended claims.

The following procedures were used to prepare several different types ofbifunctional crosslinking agents. The following procedures aremodifications of procedures reported in U.S. Pat. No. 4,101,380 andAbuchowski et al., cited above.

Example 1

Synthesis Of Polyethylene Glycol Disuccinimidyl Succinate PEG-SS2

Polyethylene glycol, PEG, (50 g, Aldrich Chemical Company, Milwaukee,Wis., sold as 3,400 average molecular weight, GPC analysis M_(n) was2,980, M_(w), was 3,480) was dissolved in 1,2-dichloroethane (250 ml)containing succinic arthydride (14.7 g) and anhydrous pyridine (12 ml).The mixture was refluxed under nitrogen for three days. After filtrationand evaporation of the solvent, the residue was dissolved in 100 mlwater and treated with the cation exchange resin Dowex™ 50 (H⁺) (50 g)for 30 minutes. The mixture was then filtered and the Dowex™ 50 waswashed with water (50 ml 1×). The combined filtrate was washed withanhydrous diethyl ether (50 ml 2×). The PEG-disuccinate was thenextracted from the water phase with two 100 ml chloroform washes.Evaporation of chloroform yielded about 49 g of PEG-disuccinate.

The PEG-disuccinate was dissolved in 200 ml N,N-dimethylformamide (DMF)at 37° C. and 4.23 g of N-hydroxysuccinimide (NHS) were added to thesolution. The mixture was cooled to 0° C. 7.58 g ofdicyclohexylcarbodiimide (DCC) were dissolved in 50 ml DMF and addeddropwise to the above solution with continuous stirring. The mixture wasleft at room temperature for 24 hours and filtered. 100 ml of toluenewere added to the filtrate and the solution was placed in an ice bath.The desired polyethylene glycol disuccinimidyl succinate product,PEG-SS2, was precipitated by slowly adding petroleum ether. Theprecipitate was collected on a 10-20 micron sintered glass filter.Dissolution in toluene and precipitation with petroleum ether wasrepeated three times. The PEG-SS2 was further purified by dissolving in100 ml of 0.1M pH 2.2 citrate/phosphate buffer and filtering through a4-8 micron sintered glass filter. The PEG-SS2 was extracted withchloroform (100 ml 2×) and the solvent was evaporated under reducedpressure in a rotary evaporator. The PEG-SS2 was then dissolved intoluene and precipitated with petroleum ether, dried under vacuumovernight at room temperature, and stored in a refrigerator.

Example 2

Synthesis of N-hydroxysuccinimide Ester of Dicarboxymethyl PolyethyleneGlycol

Dicarboxymethyl poly(ethylene glycol) (mol. wt. 3400) purchased fromShearwater Polymers, Inc., Huntsville, Ala. (5 g) andN-hydroxysuccinimide purchased from Sigma Chemical Co., St. Louis, Mo.(1 g) were dissolved in 30 ml of anhydrous DMF with mechanical stirringunder nitrogen. The solution was cooled to 0° C. and a solution ofdicyclohexylcarbodiimide (1.79 g) in 5 ml DMF was added dropwise. Thestirring was continued in the cold for 3 hours then at room temperatureovernight (16 hrs). Dicyclohexylurea which precipitated was removed byfiltration. Toluene (100 ml) was added to the filtrate and cooled to 0°C. The product was then precipitated by addition of petroleum ether. Theprecipitate was collected on a sintered glass filter. Dissolution intoluene and reprecipitation with petroleum ether was repeated threetimes. The product was dried under vacuum in a desiccator.

Example 3

Synthesis of Polyethylene Glycol-di-oligoglycolide DisuccinimidylSuccinate

A 500 ml three neck round bottom flask was flame dried under nitrogen.50 g of PEG (mol. wt. 3400), 300 ml of xylene, and 1 drop of 0.33Mstannous ottoate solution in xylene were charged into the flask with acontinuous nitrogen purge. The flask was heated to boil the solution and50 ml of xylene were removed by distillation. The solution was thencooled to room temperature. 17 g of glycolide (Boehfinger Ingleheim KG,Ingleheim, Germany) was added to the flask and the reaction mixture wasrefluxed under nitrogen for 16 hours. The copolymer reaction mixture wasfiltered hot to remove polyglycolide homopolymer. The copolymer thenprecipitated from the filtrate upon cooling and collected by filtration.The copolymer was placed in a flask with 500 ml of dichloromethane and 7g of succinyl chloride. The solution was refluxed under nitrogenovernight (16 hours). 8.5 g of N-hydroxysuccinimide was added to theflask and refluxing was continued for another overnight period. A whitesolid was obtained by precipitation upon cooling the solution. Theproduct was then purified by redissolving in toluene and reprecipitatingwith petroleum ether several times. The final precipitate was driedunder vacuum and stored in a desiccator. The structure of the productwas confirmed by NMR analysis.

Example 4

Synthesis of Polyethylene Glycol-dimaleimidyl Succinate

About 12 g of PEG-disuccinate and 1 g N-hydroxymaleimide (AldrichChemical Co.) were placed in a 250 ml three neck round bottom flask with50 ml of anhydrous DMF under nitrogen. The mixture was dissolved at 60°C. with mechanical stirring and cooled to 0° C. A solution of 1.82 gdicyclohexylcarbodiimide in DMF (5 ml) was added dropwise to the flask.The reaction was allowed to mix overnight under nitrogen at roomtemperature. Dicyclohexylurea was removed by filtration and the productwas obtained by adding toluene and precipitating with petroleum ether.Dissolution in toluene and reprecipitation with petroleum ether wererepeated three times. The purified product was dried under vacuum andstored in a desiccator.

Example 5

Synthesis of Polyethylene Glycol-diphthalimidyl Succinate

About 15 g of PEG-disuccinate and 1.65 g N-hydroxyphthalimide (AldrichChemical Co.) were placed in a 250 ml three neck round bottom flask with30 ml of anhydrous DMF under nitrogen. The mixture was dissolved at 60°C. with mechanical stirring and cooled to 0° C. A solution of 1.82 gdicyclohexylcarbodiimide in DMF (5 ml) was added dropwise to the flask.The reaction was allowed to mix overnight under nitrogen at roomtemperature. Dicyclohexylurea was removed by filtration and the productwas obtained by adding toluene and precipitating with petroleum ether.Dissolution in toluene and reprecipitation with petroleum ether wererepeated three times. The purified product was dried under vacuum andstored in a desiccator.

Example 6

Preparation of Two Component Adhesive

The following procedure was used to prepare a two-component adhesiveusing a variety of protein sources, and bifunctional crosslinkingagents. Aqueous solutions of a protein and a crosslinking agent aslisted in Table 1 were pipetted (0.2 ml of each solution) into aporcelain test well and mixed continuously with a stainless steel rod.The cure time and physical consistency of each of the two componentadhesives are also listed in Table 1.

The data indicated that fish and bovine gelatin, egg and serum albuminas well as casein protein crosslinked with PEG-SS2 provided an adhesivewhich was very elastic, had good adhesive strength and a relativelyrapid cure rate.

TABLE 1 Bifunctional Cure Protein Crosslinking agent Time ConsistencyFish Gelatin 130 mg/ml 40 sec Strong gel, very Lot 23H0307 PEG-SS2 3400mw elastic, slightly Sigma sticky 40% 0.1 M pH 10 Carb/Bicarb FishGelatin 260 mg/ml 40 sec Strong gel, very Lot 23H0307 PEG-SS2 3400 mwelastic, slightly Sigma sticky 40 % 0.1 M pH 10 Carb/Bicarb Fish Gelatin130 mg/ml 120 sec Soft gel, very Lot 23H0307 PEG-SS2 10,000 mw stickySigma 40% 0.1 M pH 10 Carb/Bicarb Fish Gelatin 260 mg/ml 110 sec Softgel to Lot 23H0307 PEG-SS2 10,000 mw elastic, Sigma moderately 40% 0.1 MpH 10 sticky Carb/Bicarb Gelatin Bovine 130 mg/ml 40 sec Soft gel, notSkin Lot 53H0271 PEG-SS2 3400 mw elastic Sigma 40% 0.1 M pH 10Carb/Bicarb Gelatin Bovine 260 mg/ml 40 sec Soft gel, not Skin Lot53H0271 PEG-SS2 3400 mw elastic Sigma 40% 0.1 M pH 10 Carb/BicarbGelatin Bovine 130 mg/ml 40 sec Soft gel, not Skin Lot 53H0271 PEG-SS210,000 mw elastic Sigma 40% 0.1 M pH 10 Carb/Bicarb Gelatin Bovine 260mg/ml 120 sec Soft gel, not Skin Lot 53H0271 PEG-SS2 10,000 mw elasticSigma 40% 0.1 M pH 10 Carb/Bicarb Casein 130 mg/ml 40 sec Strong gel, pH9.4 12.6% PEG-SS2 3400 mw elastic, not sticky Carb/Bicarb Poly-L-Lysine130 mg/ml 20 sec Waxy, no 50 mg/ml H₂O PEG-SS2 3400 mw adhesive strength300,000 mw Carb/Bicarb Poly-L-Lysine 260 mg/ml 15 sec Waxy, no 50 mg/mlH₂O PEG-SS2 3400 mw adhesive strength 300,000 mw Carb/BicarbPoly-L-Lysine 130 mg/ml 10 sec Waxy, no 50 mg/ml H₂O PEG-SS2 10,000 mwadhesive strength 300,000 mw Carb/Bicarb Poly-L-Lysine 260 mg/ml 10 secWaxy, no 50 mg/ml H₂O PEG-SS2 10,000 mw adhesive strength 300,000 mwCarb/Bicarb Chicken Egg Albumin 130 mg/ml 210 sec soft, tacky 40% 0.08 MpH 10 PEG-SS2 3400 mw Carb/Bicarb Rabit Serum Albumin 130 mg/ml 20 secVery elastic, (RSA) Sigma PEG-SS2 3400 mw good adhesive Lot 19F9301strength, not 40% 0.1 M pH 10 sticky Carb/Bicarb Human Serum Albumin 130mg/ml 20 sec Very elastic, (HSA) Sigma PEG-SS2 3400 mw good adhesive Lot63H9041 strength, not 40% 0.1 M pH 10 sticky Carb/Bicarb HSA 130 mg/ml20 sec Very elastic, Sigma PEG-SS2 3400 mw good adhesive Lot 63H0941strength, not 40% 0.1 M pH 8.44 sticky Carb/Bacarb HSA 260 mg/ml 10 secVery elastic, Sigma PEG-SS2 3400 mw good adhesive Lot 63H9041 strength,not 40% 0.1 M pH 8.44 sticky Carb/Bicarb HSA 130 mg/ml 30 sec Veryelastic, Sigma PEG-SS2 10,000 mw slight adhesive Lot 63H9041 strength,very 40% 0.1 M pH 8.44 sticky Carb/Bicarb HSA 260 mg/ml 25 sec Veryelastic, Sigma PEG-SS2 10,000 mw slight adhesive Lot 63H9041 strength,very 40% 0.1 M pH 8.44 sticky Carb/Bicarb HSA 130 mg/ml 20 sec Turnedbrown Baxter Healthcare PEG-dimaleimidyl upon curing, Corp. succinatehard gel, not Lot 2837A238AA Example 4 sticky Carb/Bicarb HSA 130 mg/ml10 sec Turned red upon Baxter PEG-diphthalimidyl curing, hard gel, Lot2837A238AA succinate not sticky Carb/Bicarb Example 5 HSA 130 mg/ml 8sec Hard gel, not Baxter PEG-dicarboxymethyl sticky, no color Lot2837A238AA disuccinimidyl change Carb/Bicarb Example 2 HSA 130 mg/ml 40sec Hard gel, not Baxter PEG-dioliglycolide sticky, no color Lot2837A238AA disuccinimidyl succinate change Carb/Bicarb Example 3 HSA 130mg/ml 30 sec Hard gel, not Baxter PEG-disuccinimidyl sticky, no colorLot 2837A238AA propionate change Carb/Bicarb PEG(SPA)2 HSA 260 mg/ml 40sec Hard gel, not Baxter PEG-disuccinimidyl sticky, no color Lot2837A238AA propionate change Carb/Bicarb PEG(SPA)2 HSA 130 mg/ml 48 hrsHard gel, not Baxter PEG-dioxycarbonyl (cure) sticky, no color Lot2837A238AA imidazole change Carb/Bicarb PEG(CDI)2 HSA 130 mg/ml 140 secHard gel, not Baxter PEG-dinitrophenyl sticky, changed Lot 2837A238AAcarbonate to bright yellow Carb/Bicarb PEG(NPC)2 color HSA 260 mg/ml 140sec Hard gel, not Baxter PEG-dinitrophenyl sticky, changed Lot2837A238AA carbonate to bright yellow Carb/Bicarb PEG(NPC)2 color HSA130 mg/ml 8 hrs Hard gel, not Baxter PEG-ditresylate (viscous) sticky,no color Lot 2837A238AA PEG(tres)2 24 hrs change Carb/Bicarb (cure) HSA130 mg/ml 72 hrs Hard gel, not Baxter PEG-diglycidyl ether (cure)sticky, no color Lot 2837A238AA PEG(epox)2 change Carb/Bicarb HSA 130mg/ml no cure Liquid Baxter PEG-dialdehyde Lot 2837A238AA PEG(ald)2Carb/Bicarb mw = weight average molecular weight

Example 7

Effect of Buffer and pH

Two component adhesives were prepared according to the process describedin Example 6 except that the pH of the buffer in the protein solutionwas changed as listed in Table 2. The data indicate that a preferred pHrange is about 8.44-10.0.

TABLE 2 Crosslinking agent Cure Protein PEG-SS2 Time Consistency HSA 130mg/ml 10 min Initially softer Baxter 3400 mw adhesive, hardness Lot2837A238AA with aging 40% 0.1 M pH 7.4 Carb/Bicarb HSA 130 mg/ml 20 secVery elastic, good Sigma 3400 mw adhesive strength, Lot 63H9041 notsticky 40% 0.1 M pH 8.44 Carb/Bicarb HSA 130 mg/ml 10 sec Hard gel, nosticky Sigma 3400 mw Lot 63H9041 40% 0.15 M pH 9.07 Carb/Bicarb HSA 130mg/ml 5 sec Hard gel, no sticky Sigma 3400 mw Lot 63H9041 40% 0.2 M pH9.52 Carb/Bicarb HSA 260 mg/ml 5 sec Hard gel, no sticky Sigma 3400 mwLot 63H9041 40% 0.2 M pH 9.52 Carb/Bicarb HSA 130 mg/ml 7 sec Elastic tohard gel, Sigma 10,000 mw slightly sticky Lot 63H9041 40% 0.2 M pH 9.52Carb/Bicarb HSA 260 mg/ml 7 sec Elastic to hard gel, Sigma 10,000 mwslightly sticky Lot 63H9041 40% 0.2 M pH 9.52 Carb/Bicarb HSA 130 mg/ml25 sec Very elastic, not Baxter 3400 mw sticky Lot 2837A238AA 40% 0.1 MpH 10 Carb/Bicarb HSA 130 mg/ml 25 sec Very elastic, not Sigma 3400 mwsticky Lot 63H9041 40% 0.1 M pH 10 Carb/Bicarb mw = weight averagemolecular weight

Example 8

Effect of Crosslinking Agent on Adhesive Strength

A 30% HSA (Human Serum Albumin) solution from Sigma Chemical Co. and a25% HSA solution from Baxter Healthcare, Inc. were dialyzed against 0.1Mcarbonate/bicarbonate pH 10 buffer at 4° C. overnight and concentratedto about 40% by ultra-filtration through a 50,000 molecular weightcut-off cellulose ester disc membrane (Spectrum Medical Industries,Inc.) in a pressure filtration cell under nitrogen at 60 psig. The finalconcentration was calculated based on the volume of collected filtrate.The maximum concentration obtained under these conditions duringovernight ultra-filtration was typically 42-45%. The RSA (Rabbit SerumAlbumin) from Sigma and RSA crystallized protein from ICN Biomedical,Inc. were dissolved in 0.1M pH 10 carbonate/bicarbonate buffer andconcentrated to 40% by the same method used for HSA.

Various concentrations of PEG-SS2 (3,400 mw and 10,000 mw) were preparedin deionized water. The albumins and crosslinking agent solutions weredelivered in equal volume using a 1 ml dual syringe. The syringe tipswere fitted with a Y connector which connected to a specially machinedTEFLQN adaptor inserted into a 1.8 in.×0.187 in. (4.57 cm×0.475 cm) dia.spiral mixer nozzle (TAH Industries, Inc., Robbinsville, N.J., part no.150-312). The adhesive mixture was injected through the mixer directlyonto the test substrate for adhesion testing.

Freshly excised guinea pig skin was cut into strips and a polystyrenewindow with an opening of 0.5×1.0 inches (1.27 cm×2.54 cm) was placed onone end of the strip to contain the glue in a specific region. Uponfilling the window with glue it was covered with another strip of guineapig skin. A 500 g steel weight was placed on top of this assembly forabout one minute. The sample was peeled apart in the jaws of a computercontrolled mechanical testing machine (880 Material Test System, MTSSystem, Inc., Minneapolis, Minn.) set at a strain rate of 0.8 in./min.(2 cm/min.) with a gage length of 1 in. (2.54 cm) and a 5 lbs. (2.27 kg)load cell. Peel force was recorded after the initiation of adhesivefailure as the constant force require to continue peeling as shown inFIG. 1. Four replicates were performed for each test condition. Theresults of this test are listed in FIG. 2.

Example 9

Measurement of Adhesive Sealant Burst Strength

A pressurization assembly illustrated in FIG. 3 was used to test thebursting strength of materials used to seal standardized holes or slitsin test membranes. This assembly included an aluminum pressure vessel(1) having a 35 mm inside diameter fitted with a millivolt output typepressure transducer (2) with a range of 0 to 15 psig (MODEL PX236, OmegaEngineering, Inc., Stamford, Conn.) and a pressure inlet port (3). Toperform a test, about a 5 mm diameter hole (4) (or other standardizeddefect) was cut in the center of a test membrane (5) using a die cutter.The membrane was then placed on a piece of 0.4 mm thick TEFLON film withthe hole in the membrane centered in a larger (14 mm diameter) hole inthe TEFLON film. The TEFLON film was then placed on a flat surface withthe membrane side down and adhesive sealant test material was applied tofill the hole in the film. A solid TEFLON block was then quickly placedover the sealant prior to cure so that the TEFLON film served as aspacer to create a layer of sealant exactly 0.4 mm thick. After thedesired cure time elapsed, the TEFLON block was inverted and themembrane was carefully peeled off to obtain a circular patch of sealant(6) covering the hole in the membrane. The test membrane was sealeddefect was then mounted onto the open end of the pressure vessel (7) byplacing it between two rubber washers (8) and then between two metalwashers (9). An air tight seal was obtained by screwing the threadedcover (10) onto the matching threads (11) of the pressure vessel. Theopening in the cover (12) was also 35 mm in diameter which, incombination with the 35 mm inside diameter washers, provided a fixedmembrane surface area for pressure testing.

Two types of membranes were used, either a collagen membrane or afreshly excised porcine pericardium sample. The porcine pericardiumsample was either used immediately upon harvest or after storage in amoisture-proof container at 4° C. for no longer than 24 hours. Underthese conditions there was no discernible difference in sealantperformance based on storage time of that tissue.

The pressurization sequence was initiated by injecting air into thepressure inlet at a fixed rate of one cubic centimeter per second usinga syringe pump (Sage Instruments Model 351, Orion Research, Inc.). Thepressure transducer was connected to a digital strain gauge meter (OmegaModel DP205-S, Omega Engineering, Inc.) programmed to read pressure (rammercury) and to display the peak pressure value at the time of adhesivesealant rupture. Replicate tests gave reproducible peak pressure valuesand the standard deviation was reported in each case.

Pressure tests were performed with an adhesive composition of 40% HSA(or RSA) in 0.08M carbonate/bicarbonate buffer at different pH valueswith 3,400 m. wt. PEG-SS2 (130 mg/ml) on collagen and pericardiummembranes. The results listed in Table 3 demonstrate excellent sealantperformance with typical peak pressure values of about 130 mm Hg.

In addition, the peak pressure for the above sealants after soaking insaline solution was measured. The test was performed as described aboveexcept that the surface of the sealant coated membrane was flooded withsaline for up to a time period of 90 minutes before pressurization.Although the sealant hydrogel swelled to about double in thickness,substantial retention of sealant performance was retained.

Table 4 shows the data obtained by testing a variety of proteinsincluding fish skin gelatin, chicken egg albumin, and fibrinogen.Fibrinogen mixed with thrombin (“fibrin glue”, BERIPLAST-P sealant,Behringwerke, Marburg, Germany) was also used as a control sealantmaterial. None of these materials performed as well as the serum albuminexamples. The main disadvantage was the cure and aging time required toachieve significant strength. In particular, chicken egg albuminrequired twenty-five minutes of post cure aging to achieve the sameburst strength obtained from serum albumin aged for less than fiveminutes.

The same process was repeated for additional 25% HSA solutions bydialyzing against 0.08M carbonate/bicarbonate buffers at pH 9 and pH 8.A pH 7 solution of HSA was obtained by concentration of the original 25%HSA solution to 40% by ultrafiltration. The crosslinking agent solutionPEG-SS2 (3400 mw) was 130 mg dissolved in one ml deionized water. Thealbumin and crosslinking agent solutions were delivered in equal volumeusing a one ml dual syringe as in Example 8. The pressure tests wereperformed as above using collagen membrane except that the sealanthydrogel was aged before testing. The results are also listed in Table4. These data demonstrate that optimal pressure test values are achievedfaster with increasing pH of the albumin solution. Moreover, theresultant cured sealant obtained after complete curing has taken placeis unexpectedly higher with higher pH of the albumin solution.

TABLE 3 Burst Adhesive Pressure Tissue Tissue Opening Composition (mmHg) Collagen 4.56 mm dia, hole HSA:PEG-SS2 150 Collagen 5 mm slitHSA:PEG-SS2 112 Collagen 4.56 mm dia, hole RSA:PEG-SS2 130 Collagen 5 mmslit RSA:PEG-SS2 125 Porcine 4.56 mm dia, hole HSA:PEG-SS2 155Pericardium Porcine 5 mm slit HSA:PEG-SS2 130 Pericardium Porcine 4.56mm dia, hole RSA:PEG-SS2 125 Pericardium Porcine 5 mm slit RSA:PEG-SS2130 Pericardium

TABLE 4 Pressure Test of Different Proteins Using Collagen andPericardium HSA: 40% 0.08 M Carb/Bicarb Buffer in Saline Lot #287a328AARSA: 40% 0.08 M Carb/Bicarb Buffer in Saline Lot #82-451-0050 INCPEG-SS2: 3400 mw lot #103128-110 (130 mg/ml) Defect: 4.5 mm hole AirFlow Rate: 1 cc/s Pressure (mm Hg) Protein Crosslinker Membrane AveStdev Comments HSA pH 10 PEG-SS2 Collagen 149 9 No bubbles Pericardium154 4 5 min after curing Pericardium 196 5 10 min after curing HSA pH 10PEG-SS2 Collagen 144 5 min after curing 155 10 min after curing 162 20min after curing HSA pH 9 PEG-SS2 Collagen 108 5 min after curing 114 10min after curing 116 20 min after curing HSA pH 8 PEG-SS2 Collagen 36 5min after curing 78 10 min after curing 90 20 min after curing HSA pH 7PEG-SS2 Collagen 30 10 min after curing 52 20 min after curing RSA pH 10PEG-SS2 Collagen 134 5 No bubbles Pericardium 126 10 5 min after curingPericardium 194 9 10 min after curing Fish Gelatin pH 10 PEG-SS2Collagen 34 2 10 min after curing 40% (Sigma) Chicken Egg PEG-SS2Collagen 14 3 10 min after curing Albumin pH 10 151 3 45 min aftercuring 40% (Sigma) Fibrin Glue Pericardium 8 2 5 min after curing(BERIPLAST-P) with saline, glue Used according to slid off easily mfg.instructions 39 2 5 min after curing without saline leaked underneathBovine Fobrinogen PEG-SS2 Collagen 8 2 5 min after curing pH 10 8 2 60min after curing 15% curing, glue slid (Sigma) off easily

Example 10

Use of a Two Component Adhesive Sealant in General and Thoracic Surgery

An anesthetized pig was used as an experimental model for thoracicsurgical complications such as staple line leaks during lung andbronchus resections, bronchopleural fistulas, and other conditionsresulting in pneumothorax.

The two component adhesive included Part A, a 40% HSA prepared bydialysis of commercially available HSA (25% Solution, BUMINATE 25%,Baxter Healthcare Corp., Hyland Division, Glendale, Calif.) against0.08M pH 10 carbonate/bicarbonate buffer followed by concentration to40% by ultrafiltration at 50 psi using a 50,000 molecular weight cut-offcellulose ester disc membrane and Part B, a 130 mg/ml solution of 3,400m.wt. PEG-SS2 dissolved in sterile distilled water no more than 30minutes prior to use. The PEG-SS2 was synthesized and purified asdescribed in Example 1.

A stab wound was made on the lung of an anesthetized pig with a scalpelwhich resulted in significant air leakage during inspiration asevidenced by bubbling of air through irrigation fluid administered tothe site. The wound was blotted with gauze to remove blood and fluid.The respirator was turned off and the adhesive was applied as a sealantusing a dual syringe (Behring PANTAJECT syringe, Behringwerke, Marburg,Germany) equipped with a spiral mixing tip. After a 20 second cure timeventilation was restored and the lung was again covered with irrigationfluid. No air leaks were observed.

A functional end-to-end anastomosis in pig intestine was conducted usinga standard stapling procedure. The adhesive material described above wasapplied to the staple lines. This resulted in a clear, adherent hydrogelcoating which appeared to seal the anastomotic line.

Under these conditions it was observed that anastomotic lines coatedwith the sealant were air tight whereas anastomotic lines not sealedwere not air tight.

Example 11

Use of Two Component Adhesive to Prevent Post-Surgical Adhesions

The tissue sealant hydrogel tested was a two part liquid system. Part Awas a sterile 40% (w/v) solution of human serum albumin in isotonic pH10 carbonate buffer (0.1M). Part B was a 400 mg/ml solution of 10,000molecular weight PEG-SS2 (polyethylene glycol disuccinimidyl succinate)in sterile distilled water prepared just prior to use. Solutions A and Bwere mixed in equal volumes with a dual syringe system connected to astatic mixing head (Tab Industries, Inc.).

Post-surgical adhesion prevention evaluation of this sealant formulationwas initiated in a series of ten female rabbits. A 2×2 cm area of theabdominal wall was excised down to the fascia on each side of theabdominal cavity exposed by a midline laparotomy incision. The uterinehorns were injured by scraping 20 times with a no. 10 scalpel blade.Each animal served as its own control by randomly applying test materialto only one of the abdominal wall injuries. The uterine horns were thenattached with two stitches to the abdominal wall within a fewmillimeters of the edge of the wound closest to the laparotomy incision.

Two weeks after surgery the rabbits were examined in order to evaluateand score the extent, type, and tenacity of adhesions present on theabdominal wall injury sites. These results are shown in Table 5. Therating system used to obtain these scores is shown in Table 6. Althoughtechnical difficulties were encountered as noted in Table 5, the testmaterial clearly provided an unexpected benefit in both the preventionof adhesions and a reduction in their severity without the presence of aknown active ingredient.

TABLE 5 Scoring of Adhesions Formed in Material EvaluationCharacteristic Extent Type Tenacity Animal Control Treatment ControlTreatment Control Treatment BAM 8 2 0+ 3 0+ 3 0+ BAM 9 3 1 3 1 3 1 BAM0+ 1 0+ 3 0+ 2 10 BAM 0* 0 0* 0 0* 0 11 BAM 4 4 3 3 3 3 12 BAM 2 1 3 2 32 13 BAM 1* 0 3* 0 3* 0 14 BAM 1 0** 1 0** 2 0** 15 BAM 1 0* 1 0* 2 0*16 BAM 1 0* 1 0* 2 0* 17 Average 1.5 0.7 1.8 0.9 2.1 0.8 *Uterine horntacked to abdominal wall with only one suture **Uterine horn no longersutured to abdominal wall +Fascia removed with peritoneum and musclelayers

TABLE 6 Adhesion Scoring Characteristic Adhesion Score Extent (%sidewall involvement) None 0 ≦25 1 ≦50 2 ≦75 3 >75 4 Type None 0 Filmy,no vessles (transparent) 1 Opaque, no vessles (translucent) 2 Opaque,small vessles present grossly 3 Opaque, larger vessles present grossly 4Tenacity None 0 Adhesions essentially fell apart 1 Adhesion lysed withtransaction 2 Adhesion required sharp dissection for lysis 3

1. An adhesive composition consisting essentially of i) a first aqueousmixture of about 20-60 wt/vol % serum albumin in about 0.01-0.25 molarbuffer at a pH in a range of about 8.0-11.0, ii) a second aqueousmixture of about 50-800 mg/ml of a crosslinking agent having a molecularweight in a range of about 1,000-15,000, wherein the crosslinking agentis of the formulaG—LM—PEG—LM—G wherein —PEG— is a diradical fragment represented by theformula—O—(CH₂—CH₂—O—)_(a)— where a is an integer from 20-300; wherein —LM— isa diradical fragment selected from the group consisting of a carbonatediradical of the formula, —C(O)—, a monoester diradical of the formula,—(CH₂)_(b)C(O)— where b is an integer from 1-5, a diester radical of theformula, —C(O)—(CH₂)_(c)—C(O)— where c is an integer from 2-10 and wherethe aliphatic portion of the diradical may be saturated or unsaturated,a dicarbonate diradical of the formula —C(O)—O—(CH₂)_(d)—O—C(O)— where dis an integer from 2-10, and an oligomeric diradical represented by theformulas —R—C(O)—, —R—C(O)—(CH₂)_(c)—C(O)—, or—R—C(O)—O—(CH₂)_(d)—O—C(O)— where c is an integer from 2-10, d is aninteger from 2-10, and R is a polymer or copolymer having 1-10 monomericfragments selected from the group consisting of lactide, glycolide,trimethylene carbonate, caprolactone and p-dioxanone; and wherein —G isa leaving group selected from the group consisting of succinimidyl,maleimidyl, phthalimidyl, imidazolyl, nitrophenyl or tresyl, and whereina combination of the first and second mixtures is initially liquid andthen cures on the surface of tissue to give a flexible, substantivematrix which bonds to the tissue and has a burst strength greater thanabout 10 mmHg.
 2. The adhesive mixture of claim 1 wherein the protein inthe first mixture is about 35-45 wt/vol % serum albumin.
 3. The adhesivecomposition of claim 1 wherein the serum albumin is human serum albumin.4. The adhesive composition of claim 1 wherein the buffer is 0.05-0.15molar carbonate/bicarbonate buffer at a pH of about 9.0-10.5.
 5. Theadhesive composition of claim 1 wherein the second aqueous mixture isabout 50-300 mg/ml of a crosslinking agent having a molecular weight ina range of about 1,000-5,000.
 6. The adhesive composition of claim 1wherein the ratio of a volume of the first mixture to a volume of thesecond mixture is in a range of about 1:10 to about 10:1.
 7. Theadhesive composition of claim 1 wherein —LM— is an oligomeric diradical—R—C(O)—(CH₂)_(c)—C(O)— where c is an integer from 2-10 and R is apolymer or copolymer having 1-10 monomeric fragments selected from thegroup consisting of lactide, glycolide, trimethylene carbonate,caprolactone and p-dioxanone.
 8. The adhesive composition of claim 1wherein —G is succinimidyl.
 9. An in vivo method of adhering tissuecomprising the steps of topically applying and bonding an adhesivemixture of claim 1 to the tissue.
 10. An in vivo method of sealing airleaks in pulmonary tissues comprising the steps of topically applyingand curing the adhesive mixture of claims 1 to an air leak site in thepulmonary tissue.
 11. An in vivo method to prevent post-surgicaladhesions comprising the step of topically applying and curing theadhesive mixture of claims 1 to tissue surrounding a surgical site. 12.An in vivo method to seal tissue comprising the step of topicallyapplying and bonding the adhesive mixture of claims 1 to tissue toprevent or control blood or other fluid leaks.
 13. The adhesivecomposition of claim 1 wherein the second aqueous mixture is about300-800 mg/ml of a crosslinking agent having a molecular weight in arange of about 5,000-15,000.
 14. The adhesive composition of claim 13wherein —LM— is a diester diradical of the formula —C(O)—(CH₂)₂—C(O)—.15. The adhesive mixture of claim 1 wherein —LM— is a diester diradicalof the formula, —C(O)—(CH₂)_(c)—C(O)— where c is an integer from 2-10and where the aliphatic portion of the diradical may be saturated orunsaturated.
 16. The adhesive composition of claim 15 wherein —LM— is aoligomeric diradical derived from polyglycolic acid.
 17. A method ofmaking a tissue adhesive consisting of the step of forming a mixture ofi) a first aqueous mixture of about 20-60 wt/vol % serum albumin inabout 0.01-0.25 molar buffer at a pH in a range of about 8.0-11.0, ii) asecond aqueous mixture of about 50-800 mg/ml of a crosslinking agenthaving a molecular weight in a range of about 1,000-15,000, wherein thecrosslinking agent is of the formula  G—LM—PEG—LM—G wherein —PEG— is adiradical fragment represented by the formula—O—(CH₂—CH₂—O—)_(a)— where a is an integer from 20-300; wherein —LM— isa diradical fragment selected from the group consisting of a carbonatediradical of the formula, —C(O)—, a monoester diradical of the formula,—(CH₂)_(b)C(O)— where b is an integer from 1-5, a diester diradical ofthe formula, —C(O)—(CH₂)_(c)—C(O)— where c is an integer from 2-10 andwhere the aliphatic portion of the diradical may be saturated orunsaturated, a dicarbonate diradical of the formula—C(O)—O—(CH₂)_(d)—O—C(O)— where d is an integer from 2-10, and anoligomeric diradical represented by the formulas —R—C(O)—,—R—C(O)—(CH₂)_(c)—C(O)—, or —R—C(O)—O—(CH₂)_(d)—O—C(O)— where c is aninteger from 2-10, d is an integer from 2-10, and R is a polymer orcopolymer having 1-10 monomeric fragments selected from the groupconsisting of lactide, glycolide, trimethylene carbonate, caprolactoneand p-dioxanone; and wherein —G is a leaving group selected from thegroup consisting of succinimidyl, maleimidyl, phthalimidyl, imidazolyl,nitrophenyl or tresyl, and wherein a combination of the first and secondmixtures is initially liquid and then cures on the surface of tissue togive a flexible, substantive matrix which bonds to the tissue and has aburst strength greater than about 10 mmHg.
 18. A method of treatingtissue to prevent or control air or fluid leaks comprising: providing acomposition to tissue, said composition including an albumin protein anda crosslinking agent, said crosslinking agent having a polyoxyethylenechain portion and an activated leaving group which allows thecrosslinking agent to react with said protein; and curing saidcomposition on the tissue to bond said composition to the tissue and toprovide a substantive cured matrix.
 19. The method of claim 18 whereinsaid composition is cured to produce the matrix in less than about 10minutes.
 20. The method of claim 18 wherein said composition is cured toproduce the matrix in less than about one minute.
 21. The method ofclaim 18 wherein said composition is cured to produce the matrix inabout ten seconds.
 22. The method of claim 18 comprising providing thecomposition to the tissue using a syringe.
 23. The method of claim 18comprising providing the composition to the tissue using a dual syringe.24. The method of claim 18 comprising providing the composition to thetissue using a spray apparatus.
 25. The method of claim 18 wherein thematrix is resorbed.
 26. The method of claim 25 wherein the matrix isresorbed in about four to sixty days.
 27. The method of claim 18comprising curing the composition such that the peel strength of thematrix is about 0.08 lb/in or more.
 28. The method of claim 18comprising curing said composition to provide a cured matrix that has aburst pressure greater than about 10 mmHg.
 29. The method of claim 27wherein the matrix has a burst pressure of about 34 mmHg or greater. 30.The method of claim 29 wherein the matrix has a burst pressure of about90 mmHg or greater.
 31. The method of claim 30 wherein the matrix has aburst pressure of about 130 mmHg or greater.
 32. The method of claim 18comprising providing a composition wherein the crosslinking agent has amolecular weight in a range of about 1,000-15,000.
 33. The method ofclaim 32 comprising providing a composition wherein the crosslinkingagent has a molecular weight in a range of about 1,000-5,000.
 34. Themethod of claim 18 comprising providing a composition wherein theactivated leaving group is an N-hydroxy imide.
 35. The method of claim34 comprising providing a composition wherein the activated leavinggroup is N-hydroxy succinimide.
 36. The method of claim 18 furthercomprising mixing a first mixture and a second mixture to form thecomposition and applying said composition to the tissue, wherein thefirst mixture includes about 20-60 wt/vol % of the protein in about0.01-0.25 molar buffer at a pH in a range of about 8.0-11.0 and thesecond mixture includes about 50-800 mg/ml of the crosslinking agenthaving a molecular weight in a range of about 1,000-15,000.
 37. Themethod of claim 36 wherein the crosslinking agent is of the formulaG—LM—PEG—LM—G wherein: —PEG— is a diradical fragment represented by theformula  —O—(CH ₂ —CH ₂ —O—)_(a) — where a is an integer from 20-300;—LM— is a diradical fragment selected from the group consisting of acarbonate diradical of the formula —C(O)—, a monoester diradical of theformula —(CH ₂)_(b) C(O)— where b is an integer from 1-5, a diesterdiradical of the formula —C(O)—(CH ₂)_(c) —C(O)— where c is an integerfrom 2-10 and where the aliphatic portion of the diradical may besaturated or unsaturated, a dicarbonate diradical of the formula—C(O)—O—(CH ₂)_(d) —O—C(O)— where d is an integer from 2-10, and anoligomeric diradical represented by the formulas —R—C(O)—, —R—C(O)—(CH₂)_(c) —C(O)—, or —R—C(O)—O—(CH ₂)_(d) —O—C(O)— where c is an integerfrom 2-10, d is an integer from 2-10, and R is a polymer or copolymerhaving 1-10 monomeric fragments selected from the group consisting oflactide, glycolide, trimethylene carbonate, caprolactone, andp-dioxanone; and —G is the leaving group selected from the groupconsisting of succinimidyl, maleimidyl, phthalimidyl, imidazolyl,nitrophenyl, and tresyl.
 38. The method of claim 37 wherein the proteinin the first mixture is about 35-45 wt/vol % serum albumin.
 39. Themethod of claim 38 wherein the buffer is 0.05-0.15 molarcarbonate/bicarbonate buffer at a pH of about 9.0-10.5.
 40. The methodof claim 37 wherein the second mixture is about 50-300 mg/ml of thecrosslinking agent having a molecular weight in a range of about1,000-5,000.
 41. The method of claim 37 wherein the ratio of a volume ofthe first mixture to a volume of the second mixture is in a range ofabout 1:10 to about 10:1.
 42. The method of claim 37 wherein —LM— is anoligomeric diradical —R—C(O)—(CH ₂)_(c) —C(O)— where c is an integerfrom 2-10 and R is a polymer or copolymer having 1-10 monomericfragments selected from the group consisting of lactide, glycolide,trimethylene carbonate, caprolactone, and p-dioxanone.
 43. The method ofclaim 37 wherein —G is succinimidyl.
 44. The method of claim 37 whereinthe second mixture includes about 300-800 mg/ml of a crosslinking agenthaving a molecular weight in a range of about 5,000-15,000.
 45. Themethod of claim 37 wherein —LM— is a diester diradical of the formula—C(O)—(CH ₂)₂ —C(O)—.
 46. The method of claim 37 wherein —LM— is adiester diradical of the formula —C(O)—(CH ₂)_(c) —C(O)— where c is aninteger from 2-10 and where the aliphatic portion of the diradical maybe saturated or unsaturated.
 47. The method of claim 37 wherein —LM— isan oligomeric diradical derived from polyglycolic acid.
 48. The methodof claim 18 comprising treating tissue to prevent or control a fluidleak.
 49. The method of claim 48 wherein the fluid leak is a blood leak.50. The method of claim 18 wherein the tissue includes an air leak. 51.The method of claim 50 wherein the air leak is in a pulmonary system.52. A method of treating tissue to prevent formation of an adhesioncomprising: providing a composition to tissue, said compositionincluding an albumin protein and a crosslinking agent, said crosslinkingagent having a polyoxyethylene chain portion and an activated leavinggroup which allows the crosslinking agent to react with said protein;and curing said composition on the tissue to bond said composition tothe tissue and to provide a substantive cured matrix.
 53. The method ofclaim 52 wherein said composition is cured to produce the matrix in lessthan about 10 minutes.
 54. The method of claim 52 wherein saidcomposition is cured to produce the matrix in less than about oneminute.
 55. The method of claim 52 wherein said composition is cured toproduce the matrix in about ten seconds.
 56. The method of claim 52comprising providing the composition to the tissue using a syringe. 57.The method of claim 52 comprising providing the composition to thetissue using a dual syringe.
 58. The method of claim 52 comprisingproviding the composition to the tissue using a spray apparatus.
 59. Themethod of claim 52 wherein the matrix is resorbed.
 60. The method ofclaim 59 wherein the matrix is resorbed in about four to sixty days. 61.The method of claim 52 comprising curing the composition such that thepeel strength of the matrix is about 0.08 lb/in or more.
 62. The methodof claim 52 comprising curing said composition to provide a cured matrixthat has a burst pressure greater than about 10 mmHg.
 63. The method ofclaim 62 wherein the matrix has a burst pressure of about 34 mmHg orgreater.
 64. The method of claim 63 wherein the matrix has a burstpressure of about 90 mmHg or greater.
 65. The method of claim 64 whereinthe matrix has a burst pressure of about 130 mmHg or greater.
 66. Themethod of claim 52 comprising providing a composition wherein thecrosslinking agent has a molecular weight in a range of about1,000-15,000.
 67. The method of claim 66 comprising providing acomposition wherein the crosslinking agent has a molecular weight in arange of about 1,000-5,000.
 68. The method of claim 52 comprisingproviding a composition wherein the activated leaving group is anN-hydroxy imide.
 69. The method of claim 68 comprising providing acomposition wherein the activated leaving group is N-hydroxysuccinimide.
 70. The method of claim 52 further comprising mixing afirst mixture and a second mixture to form the composition and applyingsaid composition to the tissue, wherein the first mixture includes about20-60 wt/vol % of the protein in about 0.01-0.25 molar buffer at a pH ina range of about 8.0-11.0 and the second mixture includes about 50-800mg/ml of the crosslinking agent having a molecular weight in a range ofabout 1,000-15,000.
 71. The method of claim 70 wherein the crosslinkingagent is of the formulaG—LM—PEG—LM—G wherein: —PEG— is a diradical fragment represented by theformula—O—(CH ₂ —CH ₂ —O—)_(a) — wherein a is an integer from 20-300; —LM— is adiradical fragment selected from the group consisting of a carbonatediradical of the formula —C(O)—, a monoester diradical of the formula—(CH ₂)_(b) C(O)— where b is an integer from 1-5, a diester diradical ofthe formula —C(O)—(CH ₂)_(c) —C(O)— where c is an integer from 2-10 andwhere the aliphatic portion of the diradical may be saturated orunsaturated, a dicarbonate diradical of the formula —C(O)—O—(CH ₂)_(d)—O—C(O)— where d is an integer from 2-10, and an oligomeric diradicalrepresented by the formulas —R—C(O)—, —R—C(O)—(CH ₂)_(c) —C(O)—, or—R—C(O)—O—(CH ₂)_(d) —O—C(O)— where c is an integer from 2-10, d is aninteger from 2-10, and R is a polymer or copolymer having 1-10 monomericfragments selected from the group consisting of lactide, glycolide,trimethylene carbonate, caprolactone, and p-dioxanone; and —G is theleaving group selected from the group consisting of succinimidyl,maleimidyl, phthalimidyl, imidazolyl, nitrophenyl, and tresyl.
 72. Themethod of claim 71 wherein the protein in the first mixture is about35-45 wt/vol % serum albumin.
 73. The method of claim 72 wherein thebuffer is 0.05-0.15 molar carbonate/bicarbonate buffer at a pH of about9.0-10.5.
 74. The method of claim 71 wherein the second mixture is about50-300 mg/ml of the crosslinking agent having a molecular weight in arange of about 1,000-5,000.
 75. The method of claim 71 wherein the ratioof a volume of the first mixture to a volume of the second mixture is ina range of about 1:10 to about 10:1.
 76. The method of claim 71 wherein—LM— is an oligomeric diradical —R—C(O)—(CH ₂)_(c) —C(O)— where c is aninteger from 2-10 and R is a polymer or copolymer having 1-10 monomericfragments selected from the group consisting of lactide, glycolide,trimethylene carbonate, caprolactone, and p-dioxanone.
 77. The method ofclaim 71 wherein —G is succinimidyl.
 78. The method of claim 71 whereinthe second mixture includes about 300-800 mg/ml of a crosslinking agenthaving a molecular weight in a range of about 5,000-15,000.
 79. Themethod of claim 71 wherein —LM— is a diester diradical of the formula—C(O)—(CH ₂)₂ —C(O)—.
 80. The method of claim 71 wherein —LM— is adiester diradical of the formula —C(O)—(CH ₂)_(c) —C(O)— where c is aninteger from 2-10 and where the aliphatic portion of the diradical maybe saturated or unsaturated.
 81. The method of claim 71 wherein —LM— isan oligomeric diradical derived from polyglycolic acid.
 82. The methodof claim 52 wherein the composition is provided to tissue at a surgicalsite.
 83. The method of claim 52 wherein the composition is provided ona surface of an internal organ.
 84. A method of treating tissue to bindlayers of tissue together comprising: providing a composition to tissue,said composition including an albumin protein and a crosslinking agent,said crosslinking agent having a polyoxyethylene chain portion and anactivated leaving group which allows the crosslinking agent to reactwith said protein; and curing said composition on the tissue to bondsaid composition to the tissue and to provide a substantive curedmatrix.
 85. The method of claim 84 wherein said composition is cured toproduce the matrix in less than about 10 minutes.
 86. The method ofclaim 84 wherein said composition is cured to produce the matrix in lessthan about one minute.
 87. The method of claim 84 wherein saidcomposition is cured to produce the matrix in about ten seconds.
 88. Themethod of claim 84 comprising providing the composition to the tissueusing a syringe.
 89. The method of claim 84 comprising providing thecomposition to the tissue using a dual syringe.
 90. The method of claim84 comprising providing the composition to the tissue using a sprayapparatus.
 91. The method of claim 84 wherein the matrix is resorbed.92. The method of claim 91 wherein the matrix is resorbed in about fourto sixty days.
 93. The method of claim 84 comprising curing thecomposition such that the peel strength of the matrix is about 0.08lb/in or more.
 94. The method of claim 84 comprising curing saidcomposition to provide a cured matrix that has a burst pressure greaterthan about 10 mmHg.
 95. The method of claim 94 wherein the matrix has aburst pressure of about 34 mmHg or greater.
 96. The method of claim 95wherein the matrix has a burst pressure of about 90 mmHg or greater. 97.The method of claim 96 wherein the matrix has a burst pressure of about130 mmHg or greater.
 98. The method of claim 84 comprising providing acomposition wherein the crosslinking agent has a molecular weight in arange of about 1,000-15,000.
 99. The method of claim 98 comprisingproviding a composition wherein the crosslinking agent has a molecularweight in a range of about 1,000-5,000.
 100. The method of claim 84comprising providing a composition wherein the activated leaving groupis an N-hydroxy imide.
 101. The method of claim 100 comprising providinga composition wherein the activated leaving group is N-hydroxysuccinimide.
 102. The method of claim 84 further comprising mixing afirst mixture and a second mixture to form the composition and applyingsaid composition to the tissue, wherein the first mixture includes about20-60 wt/vol % of the protein in about 0.01-0.25 molar buffer at a pH ina range of about 8.0-11.0 and the second mixture includes about 50-800mg/ml of the crosslinking agent having a molecular weight in a range ofabout 1,000-15,000.
 103. The method of claim 102 wherein thecrosslinking agent is of the formulaG—LM—PEG—LM—G wherein: —PEG— is a diradical fragment represented by theformula—O—(CH ₂ —CH ₂ —O—)_(a) — where a is an integer from 20-300; —LM— is adiradical fragment selected from the group consisting of a carbonatediradical of the formula —C(O)—, a monoester diradical of the formula—(CH ₂)_(b) C(O)— where b is an integer from 1-5, a diester diradical ofthe formula —C(O)—(CH ₂)_(c) —C(O)— where c is an integer from 2-10 andwhere the aliphatic portion of the diradical may be saturated orunsaturated, a dicarbonate diradical of the formula —C(O)—O—(CH ₂)_(d)—O—C(O)— where d is an integer from 2-10, and an oligomeric diradicalrepresented by the formulas —R—C(O)—, —R—C(O)—(CH ₂)_(c) —C(O)—, or—R—C(O)—O—(CH ₂)_(d) —O—C(O)— where c is an integer from 2-10, d is aninteger from 2-10, and R is a polymer or copolymer having 1-10 monomericfragments selected from the group consisting of lactide, glycolide,trimethylene carbonate, caprolactone, and p-dioxanone; and —G is theleaving group selected from the group consisting of succinimidyl,maleimidyl, phthalimidyl, imidazolyl, nitrophenyl, and tresyl.
 104. Themethod of claim 103 wherein the protein in the first mixture is about35-45 wt/vol % serum albumin.
 105. The method of claim 104 wherein thebuffer is 0.05-0.15 molar carbonate/bicarbonate buffer at a pH of about9.0-10.5.
 106. The method of claim 103 wherein the second mixture isabout 50-300 mg/ml of the crosslinking agent having a molecular weightin a range of about 1,000-5,000.
 107. The method of claim 103 whereinthe ratio of a volume of the first mixture to a volume of the secondmixture is in a range of about 1:10 to about 10:1.
 108. The method ofclaim 103 wherein —LM— is an oligomeric diradical —R—C(O)—(CH ₂)_(c)—C(O)— where c is an integer from 2-10 and R is a polymer or copolymerhaving 1-10 monomeric fragments selected from the group consisting oflactide, glycolide, trimethylene carbonate, caprolactone, andp-dioxanone.
 109. The method of claim 103 wherein —G is succinimidyl.110. The method of claim 103 wherein the second mixture includes about300-800 mg/ml of a crosslinking agent having a molecular weight in arange of about 5,000-15,000.
 111. The method of claim 103 wherein —LM—is a diester diradical of the formula —C(O)—(CH ₂)₂ —C(O)—.
 112. Themethod of claim 103 wherein —LM— is a diester diradical of the formula—C(O)—(CH ₂)_(c) —C(O)— where c is an integer from 2-10 and where thealiphatic portion of the diradical may be saturated or unsaturated. 113.The method of claim 103 wherein —LM— is an oligomeric diradical derivedfrom polyglycolic acid.
 114. The method of claim 84 wherein the matrixbinds tissue together in addition to a suture, a staple, a tape, or abandage.
 115. The method of claim 84 wherein the composition is providedto attach skin grafts.
 116. The method of claim 84 wherein thecomposition is provided to attach adjacent layers of tissue.
 117. Themethod of claim 84 wherein the composition is provided to positiontissue flaps.
 118. The method of claim 84 wherein the composition isprovided to close gingival flaps.
 119. A method of treating tissuecomprising: providing a composition to tissue, said compositionincluding an albumin protein and a crosslinking agent, said crosslinkingagent having a polyoxyethylene chain portion and an activated leavinggroup which allows the crosslinking agent to react with said protein;and curing said composition on the tissue to bond said composition tothe tissue and to provide a substantive cured matrix.
 120. The method ofclaim 119 wherein said composition is cured to produce the matrix inless than about 10 minutes.
 121. The method of claim 119 wherein saidcomposition is cured to produce the matrix in less than about oneminute.
 122. The method of claim 119 wherein said composition is curedto produce the matrix in about ten seconds.
 123. The method of claim 119comprising providing the composition to the tissue using a syringe. 124.The method of claim 119 comprising providing the composition to thetissue using a dual syringe.
 125. The method of claim 119 comprisingproviding the composition to the tissue using a spray apparatus. 126.The method of claim 119 wherein the matrix is resorbed.
 127. The methodof claim 126 wherein the matrix is resorbed in about four to sixty days.128. The method of claim 119 comprising curing the composition such thatthe peel strength of the matrix is about 0.08 lb/in or more.
 129. Themethod of claim 119 comprising curing said composition to provide acured matrix that has a burst pressure greater than about 10 mmHg. 130.The method of claim 129 wherein the matrix has a burst pressure of about34 mmHg or greater.
 131. The method of claim 130 wherein the matrix hasa burst pressure of about 90 mmHg or greater.
 132. The method of claim131 wherein the matrix has a burst pressure of about 130 mmHg orgreater.
 133. The method of claim 119 comprising providing a compositionwherein the crosslinking agent has a molecular weight in a range ofabout 1,000-15,000.
 134. The method of claim 133 comprising providing acomposition wherein the crosslinking agent has a molecular weight in arange of about 1,000-5,000.
 135. The method of claim 119 comprisingproviding a composition wherein the activated leaving group is anN-hydroxy imide.
 136. The method of claim 135 comprising providing acomposition wherein the activated leaving group is N-hydroxysuccinimide.
 137. The method of claim 119 further comprising mixing afirst mixture and a second mixture to form the composition and applyingsaid composition to the tissue, wherein the first mixture includes about20-60 wt/vol % of the protein in about 0.01-0.25 molar buffer at a pH ina range of about 8.0-11.0 and the second mixture includes about 50-800mg/ml of the crosslinking agent having a molecular weight in a range ofabout 1,000-15,000.
 138. The method of claim 137 wherein thecrosslinking agent is of the formulaG—LM—PEG—LM—G wherein: —PEG— is a diradical fragment represented by theformula  —O—(CH ₂ —CH ₂ —O—)_(a) — where a is an integer from 20-300;—LM— is a diradical fragment selected from the group consisting of acarbonate diradical of the formula —C(O)—, a monoester diradical of theformula —(CH ₂)_(b) C(O)— where b is an integer from 1-5, a diesterdiradical of the formula —C(O)—(CH ₂)_(c) —C(O)— where c is an integerfrom 2-10 and where the aliphatic portion of the diradical may besaturated or unsaturated, a dicarbonate diradical of the formula—C(O)—O—(CH ₂)_(d) —O—C(O)— where d is an integer from 2-10, and anoligomeric diradical represented by the formulas —R—C(O)—, —R—C(O)—(CH₂)_(c) —C(O)—, or —R—C(O)—O—(CH ₂)_(d) —O—C(O)— where c is an integerfrom 2-10, d is an integer from 2-10, and R is a polymer or copolymerhaving 1-10 monomeric fragments selected from the group consisting oflactide, glycolide, trimethylene carbonate, caprolactone, andp-dioxanone; and —G is the leaving group selected from the groupconsisting of succinimidyl, maleimidyl, phthalimidyl, imidazolyl,nitrophenyl, and tresyl.
 139. The method of claim 138 wherein theprotein in the first mixture is about 35-45 wt/vol % serum albumin. 140.The method of claim 139 wherein the buffer is 0.05-0.15 molarcarbonate/bicarbonate buffer at a pH of about 9.0-10.5.
 141. The methodof claim 138 wherein the second mixture is about 50-300 mg/ml of thecrosslinking agent having a molecular weight in a range of about1,000-5,000.
 142. The method of claim 138 wherein the ratio of a volumeof the first mixture to a volume of the second mixture is in a range ofabout 1:10 to about 10:1.
 143. The method of claim 138 wherein —LM— isan oligomeric diradical —R—C(O)—(CH ₂)_(c) —C(O)— where c is an integerfrom 2-10 and R is a polymer or copolymer having 1-10 monomericfragments selected from the group consisting of lactide, glycolide,trimethylene carbonate, caprolactone, and p-dioxanone.
 144. The methodof claim 138 wherein —G is succinimidyl.
 145. The method of claim 138wherein the second mixture includes about 300-800 mg/ml of acrosslinking agent having a molecular weight in a range of about5,000-15,000.
 146. The method of claim 138 wherein —LM— is a diesterdiradical of the formula —C(O)—(CH ₂)₂ —C(O)—.
 147. The method of claim138 wherein —LM— is a diester diradical of the formula —C(O)—(CH ₂)_(c)—C(O)— where c is an integer from 2-10 and where the aliphatic portionof the diradical may be saturated or unsaturated.
 148. The method ofclaim 138 wherein —LM— is an oligomeric diradical derived frompolyglycolic acid.
 149. The method of claim 119 comprising curing thecomposition on the tissue to seal the tissue.
 150. The method of claim149 comprising treating tissue to prevent or control a fluid leak. 151.The method of claim 150 wherein the fluid leak is a blood leak.
 152. Themethod of claim 149 wherein the tissue includes an air leak.
 153. Themethod of claim 152 wherein the air leak is in a pulmonary system. 154.The method of claim 119 wherein the composition is provided to tissue ata surgical site.
 155. The method of claims 119 comprising curing thecomposition at the tissue to prevent a tissue adhesion.
 156. The methodof claim 119 wherein the composition is provided on a surface of aninternal organ.
 157. The method of claim 119 comprising curing thecomposition to form a matrix to bind tissue.
 158. The method of claim157 wherein the matrix binds tissue together in addition to a suture, astaple, a tape, or a bandage.
 159. The method of claim 119 wherein thecomposition is provided to attach skin grafts.
 160. The method of claim119 wherein the composition is provided to attach adjacent layers oftissue.
 161. The method of claim 119 wherein the composition is providedto position tissue flaps.
 162. The method of claim 119 wherein thecomposition is provided to close gingival flaps.
 163. A method oftreating tissue to prevent or control air or fluid leaks comprising:providing a composition to tissue, said composition including an albuminprotein at about 20-60 wt/vol % and a crosslinking agent at about 50-800mg/ml, said crosslinking agent having a polyoxyethylene chain portionand an activated leaving group which allows the crosslinking agent toreact with said protein and having a molecular weight of about1000-15,000; and curing said composition on the tissue to bond saidcomposition to the tissue and to provide a substantive cured matrix.164. The method of claim 163 wherein said composition is cured toproduce the matrix in less than about 10 minutes.
 165. The method ofclaim 163 wherein said composition is cured to produce the matrix inless than about one minute.
 166. The method of claim 163 wherein saidcomposition is cured to produce the matrix in about ten seconds. 167.The method of claim 163 comprising providing the composition to thetissue using a syringe.
 168. The method of claim 163 comprisingproviding the composition to the tissue using a dual syringe.
 169. Themethod of claim 163 comprising providing the composition to the tissueusing a spray apparatus.
 170. The method of claim 163 wherein the matrixis resorbed.
 171. The method of claim 170 wherein the matrix is resorbedin about four to sixty days.
 172. The method of claim 163 comprisingcuring the composition such that the peel strength of the matrix isabout 0.08 lb/in or more.
 173. The method of claim 163 comprising curingsaid composition to provide a cured matrix that has a burst pressuregreater than about 10 mmHg.
 174. The method of claim 173 wherein thematrix has a burst pressure of about 34 mmHg or greater.
 175. The methodof claim 174 wherein the matrix has a burst pressure of about 90 mmHg orgreater.
 176. The method of claim 175 wherein the matrix has a burstpressure of about 130 mmHg or greater.
 177. The method of claim 163comprising providing a composition wherein the crosslinking agent has amolecular weight in a range of about 1,000-5,000.
 178. The method ofclaim 163 comprising providing a composition wherein the activatedleaving group is an N-hydroxy imide.
 179. The method of claim 178comprising providing a composition wherein the activated leaving groupis N-hydroxy succinimide.
 180. The method of claim 163 furthercomprising mixing a first mixture and a second mixture to form thecomposition and applying said composition to the tissue, wherein thefirst mixture includes about 20-60 wt/vol % of the protein in about0.01-0.25 molar buffer at a pH in a range of about 8.0-11.0 and thesecond mixture includes about 50-800 mg/ml of the crosslinking agenthaving a molecular weight in a range of about 1,000-15,000.
 181. Themethod of claim 180 wherein the crosslinking agent is of the formulaG—LM—PEG—LM—G wherein: —PEG— is a diradical fragment represented by theformula—O—(CH ₂ —CH ₂ —O—)_(a) — where a is an integer from 20-300; —LM— is adiradical fragment selected from the group consisting of a carbonatediradical of the formula —C(O)—, a monoester diradical of the formula—(CH ₂)_(b) C(O)— where b is an integer from 1-5, a diester diradical ofthe formula —C(O)—(CH ₂)_(c) —C(O)— where c is an integer from 2-10 andwhere the aliphatic portion of the diradical may be saturated orunsaturated, a dicarbonate diradical of the formula —C(O)—O—(CH ₂)_(d)—O—C(O)— where d is an integer from 2-10, and an oligomeric diradicalrepresented by the formulas —R—C(O)—, —R—C(O)—(CH ₂)_(c) —C(O)—, or—R—C(O)—O—(CH ₂)_(d) —O—C(O)— where c is an integer from 2-10, d is aninteger from 2-10, and R is a polymer or copolymer having 1-10 monomericfragments selected from the group consisting of lactide, glycolide,trimethylene carbonate, caprolactone, and p-dioxanone; and —G is theleaving group selected from the group consisting of succinimidyl,maleimidyl, phthalimidyl, imidazolyl, nitrophenyl, and tresyl.
 182. Themethod of claim 181 wherein the protein in the first mixture is about35-45 wt/vol % serum albumin.
 183. The method of claim 182 wherein thebuffer is 0.05-0.15 molar carbonate/bicarbonate buffer at a pH of about9.0-10.5.
 184. The method of claim 181 wherein the second mixture isabout 50-300 mg/ml of the crosslinking agent having a molecular weightin a range of about 1,000-5,000.
 185. The method of claim 181 whereinthe ratio of a volume of the first mixture to a volume of the secondmixture is in a range of about 1:10 to about 10:1.
 186. The method ofclaim 181 wherein —LM— is an oligomeric diradical —R—C(O)—(CH ₂)_(c)—C(O)— where c is an integer from 2-10 and R is a polymer or copolymerhaving 1-10 monomeric fragments selected from the group consisting oflactide, glycolide, trimethylene carbonate, caprolactone, andp-dioxanone.
 187. The method of claim 181 wherein —G is succinimidyl.188. The method of claim 181 wherein the second mixture includes about300-800 mg/ml of a crosslinking agent having a molecular weight in arange of about 5,000-15,000.
 189. The method of claim 181 wherein —LM—is a diester diradical of the formula —C(O)—(CH ₂)₂ —C(O)—.
 190. Themethod of claim 181 wherein —LM— is a diester diradical of the formula—C(O)—(CH ₂)_(c) —C(O)— where c is an integer from 2-10 and where thealiphatic portion of the diradical may be saturated or unsaturated. 191.The method of claim 181 wherein —LM— is an oligomeric diradical derivedfrom polyglycolic acid.
 192. The method of claim 163 comprising treatingtissue to prevent or control a fluid leak.
 193. The method of claim 192wherein the fluid leak is a blood leak.
 194. The method of claim 163wherein the tissue includes an air leak.
 195. The method of claim 194wherein the air leak is in a pulmonary system.
 196. A method of treatingtissue to prevent formation of an adhesion comprising: providing acomposition to tissue, said composition including an albumin protein atabout 20-60 wt/vol % and a crosslinking agent at about 50-800 mg/ml,said crosslinking agent having a polyoxyethylene chain portion and anactivated leaving group which allows the crosslinking agent to reactwith said protein and having a molecular weight in a range of about1000-15,000; and curing said composition on the tissue to bond saidcomposition to the tissue and to provide a substantive cured matrix.197. The method of claim 196 wherein said composition is cured toproduce the matrix in less than about 10 minutes.
 198. The method ofclaim 196 wherein said composition is cured to produce the matrix inless than about one minute.
 199. The method of claim 196 wherein saidcomposition is cured to produce the matrix in about ten seconds. 200.The method of claim 196 comprising providing the composition to thetissue using a syringe.
 201. The method of claim 196 comprisingproviding the composition to the tissue using a dual syringe.
 202. Themethod of claim 196 comprising providing the composition to the tissueusing a spray apparatus.
 203. The method of claim 196 wherein the matrixis resorbed.
 204. The method of claim 203 wherein the matrix is resorbedin about four to sixty days.
 205. The method of claim 196 comprisingcuring the composition such that the peel strength of the matrix isabout 0.08 lb/in or more.
 206. The method of claim 196 comprising curingsaid composition to provide a cured matrix that has a burst pressuregreater than about 10 mmHg.
 207. The method of claim 206 wherein thematrix has a burst pressure of about 34 mmHg or greater.
 208. The methodof claim 207 wherein the matrix has a burst pressure of about 90 mmHg orgreater.
 209. The method of claim 208 wherein the matrix has a burstpressure of about 130 mmHg or greater.
 210. The method of claim 196comprising providing a composition wherein the crosslinking agent has amolecular weight in a range of about 1,000-5,000.
 211. The method ofclaim 196 comprising providing a composition wherein the activatedleaving group is an N-hydroxy imide.
 212. The method of claim 211comprising providing a composition wherein the activated leaving groupis N-hydroxy succinimide.
 213. The method of claim 196 furthercomprising mixing a first mixture and a second mixture to form thecomposition and applying said composition to the tissue, wherein thefirst mixture includes about 20-60 wt/vol % of the protein in about0.01-0.25 molar buffer at a pH in a range of about 8.0-11.0 and thesecond mixture includes about 50-800 mg/ml of the crosslinking agenthaving a molecular weight in a range of about 1,000-15,000.
 214. Themethod of claim 213 wherein the crosslinking agent is of the formulaG—LM—PEG—LM—G wherein: —PEG— is a diradical fragment represented by theformula—O—(CH ₂ —CH ₂ —O—)_(a) — where a is an integer from 20-300; —LM— is adiradical fragment selected from the group consisting of a carbonatediradical of the formula —C(O)—, a monoester diradical of the formula—(CH ₂)_(b) C(O)— where b is an integer from 1-5, a diester diradical ofthe formula —C(O)—(CH ₂)_(c) —C(O)— where c is an integer from 2-10 andwhere the aliphatic portion of the diradical may be saturated orunsaturated, a dicarbonate diradical of the formula —C(O)—O—(CH ₂)_(d)—O—C(O)— where d is an integer from 2-10, and an oligomeric diradicalrepresented by the formulas —R—C(O)—, —R—C(O)—(CH ₂)_(c) —C(O)—, or—R—C(O)—O—(CH ₂)_(d) —O—C(O)— where c is an integer from 2-10, d is aninteger from 2-10, and R is a polymer or copolymer having 1-10 monomericfragments selected from the group consisting of lactide, glycolide,trimethylene carbonate, caprolactone, and p-dioxanone; and —G is theleaving group selected from the group consisting of succinimidyl,maleimidyl, phthalimidyl, imidazolyl, nitrophenyl, and tresyl.
 215. Themethod of claim 214 wherein the protein in the first mixture is about35-45 wt/vol % serum albumin.
 216. The method of claim 215 wherein thebuffer is 0.05-0.15 molar carbonate/bicarbonate buffer at a pH of about9.0-10.5.
 217. The method of claim 214 wherein the second mixture isabout 50-300 mg/ml of the crosslinking agent having a molecular weightin a range of about 1,000-5,000.
 218. The method of claim 214 whereinthe ratio of a volume of the first mixture to a volume of the secondmixture is in a range of about 1:10 to about 10:1.
 219. The method ofclaim 214 wherein —LM— is an oligomeric diradical —R—C(O)—(CH ₂)_(c)—C(O)— where c is an integer from 2-10 and R is a polymer or copolymerhaving 1-10 monomeric fragments selected from the group consisting oflactide, glycolide, trimethylene carbonate, caprolactone, andp-dioxanone.
 220. The method of claim 214 wherein —G is succinimidyl.221. The method of claim 214 wherein the second mixture includes about300-800 mg/ml of a crosslinking agent having a molecular weight in arange of about 5,000-15,000.
 222. The method of claim 214 wherein —LM—is a diester diradical of the formula —C(O)—(CH ₂)₂ —C(O)—.
 223. Themethod of claim 214 wherein —LM— is a diester diradical of the formula—C(O)—(CH ₂)_(c) —C(O)— where c is an integer from 2-10 and where thealiphatic portion of the diradical may be saturated or unsaturated. 224.The method of claim 214 wherein —LM— is an oligomeric diradical derivedfrom polyglycolic acid.
 225. The method of claim 196 wherein thecomposition is provided to tissue at a surgical site.
 226. The method ofclaim 196 wherein the composition is provided on a surface of aninternal organ.
 227. A method of treating tissue to bind layers oftissue together comprising: providing a composition to tissue, saidcomposition including an albumin protein of about 20-60 wt/vol % and acrosslinking agent at about 50-800 mg/ml, said crosslinking agent havinga polyoxyethylene chain portion and an activated leaving group whichallows the crosslinking agent to react with said protein having amolecular weight in a range of about 1000-15,000; and curing saidcomposition on the tissue to bond said composition to the tissue and toprovide a substantive cured matrix.
 228. The method of claim 227 whereinsaid composition is cured to produce the matrix in less than about 10minutes.
 229. The method of claim 227 wherein said composition is curedto produce the matrix in less than about one minute.
 230. The method ofclaim 227 wherein said composition is cured to produce the matrix inabout ten seconds.
 231. The method of claim 227 comprising providing thecomposition to the tissue using a syringe.
 232. The method of claim 227comprising providing the composition to the tissue using a dual syringe.233. The method of claim 227 comprising providing the composition to thetissue using a spray apparatus.
 234. The method of claim 227 wherein thematrix is resorbed.
 235. The method of claim 234 wherein the matrix isresorbed in about four to sixty days.
 236. The method of claim 227comprising curing the composition such that the peel strength of thematrix is about 0.08 lb/in or more.
 237. The method of claim 227comprising curing said composition to provide a cured matrix that has aburst pressure greater than about 10 mmHg.
 238. The method of claim 237wherein the matrix has a burst pressure of about 34 mmHg or greater.239. The method of claim 238 wherein the matrix has a burst pressure ofabout 90 mmHg or greater.
 240. The method of claim 239 wherein thematrix has a burst pressure of about 130 mmHg or greater.
 241. Themethod of claim 227 comprising providing a composition wherein thecrosslinking agent has a molecular weight in a range of about1,000-5,000.
 242. The method of claim 227 comprising providing acomposition wherein the activated leaving group is an N-hydroxy imide.243. The method of claim 242 comprising providing a composition whereinthe activated leaving group is N-hydroxy succinimide.
 244. The method ofclaim 227 further comprising mixing a first mixture and a second mixtureto form the composition and applying said composition to the tissue,wherein the first mixture includes about 20-60 wt/vol % of the proteinin about 0.01-0.25 molar buffer at a pH in a range of about 8.0-11.0 andthe second mixture includes about 50-800 mg/ml of the crosslinking agenthaving a molecular weight in a range of about 1,000-15,000.
 245. Themethod of claim 244 wherein the crosslinking agent is of the formulaG—LM—PEG—LM—G wherein: —PEG— is a diradical fragment represented by theformula—O—(CH ₂ —CH ₂ —O—)_(a) — where a is an integer from 20-300; —LM— is adiradical fragment selected from the group consisting of a carbonatediradical of the formula —C(O)—, a monoester diradical of the formula—(CH ₂)_(b) C(O)— where b is an integer from 1-5, a diester diradical ofthe formula —C(O)—(CH ₂)_(c) —C(O)— where c is an integer from 2-10 andwhere the aliphatic portion of the diradical may be saturated orunsaturated, a dicarbonate diradical of the formula —C(O)—O—(CH ₂)_(d)—O—C(O)— where d is an integer from 2-10, and an oligomeric diradicalrepresented by the formulas —R—C(O)—, —R—C(O)—(CH ₂)_(c) —C(O)—, or—R—C(O)—O—(CH ₂)_(d) —O—C(O)— where c is an integer from 2-10, d is aninteger from 2-10, and R is a polymer or copolymer having 1-10 monomericfragments selected from the group consisting of lactide, glycolide,trimethylene carbonate, caprolactone, and p-dioxanone; and —G is theleaving group selected from the group consisting of succinimidyl,maleimidyl, phthalimidyl, imidazolyl, nitrophenyl, and tresyl.
 246. Themethod of claim 245 wherein the protein in the first mixture is about35-45 wt/vol % serum albumin.
 247. The method of claim 246 wherein thebuffer is 0.05-0.15 molar carbonate/bicarbonate buffer at a pH of about9.0-10.5.
 248. The method of claim 245 wherein the second mixture isabout 50-300 mg/ml of the crosslinking agent having a molecular weightin a range of about 1,000-5,000.
 249. The method of claim 245 whereinthe ratio of a volume of the first mixture to a volume of the secondmixture is in a range of about 1:10 to about 10:1.
 250. The method ofclaim 245 wherein —LM— is an oligomeric diradical —R—C(O)—(CH ₂)_(c)—C(O)— where c is an integer from 2-10 and R is a polymer or copolymerhaving 1-10 monomeric fragments selected from the group consisting oflactide, glycolide, trimethylene carbonate, caprolactone, andp-dioxanone.
 251. The method of claim 245 wherein —G is succinimidyl.252. The method of claim 245 wherein the second mixture includes about300-800 mg/ml of a crosslinking agent having a molecular weight in arange of about 5,000-15,000.
 253. The method of claim 245 wherein —LM—is a diester diradical of the formula —C(O)—(CH ₂)₂ —C(O)—.
 254. Themethod of claim 245 wherein —LM— is a diester diradical of the formula—C(O)—(CH ₂)_(c) —C(O)— where c is an integer from 2-10 and where thealiphatic portion of the diradical may be saturated or unsaturated. 255.The method of claim 245 wherein —LM— is an oligomeric diradical derivedfrom polyglycolic acid.
 256. The method of claim 227 wherein the matrixbinds tissue together in addition to a suture, a staple, a tape, or abandage.
 257. The method of claim 227 wherein the composition isprovided to attach skin grafts.
 258. The method of claim 227 wherein thecomposition is provided to attach adjacent layers of tissue.
 259. Themethod of claim 227 wherein the composition is provided to positiontissue flaps.
 260. The method of claim 227 wherein the composition isprovided to close gingival flaps.
 261. A method of treating tissuecomprising: providing a composition to tissue, said compositionincluding an albumin protein and a crosslinking agent at about 20-60wt/vol %, said crosslinking agent of about 50-800 mg/ml having apolyoxyethylene chain portion and an activated leaving group whichallows the crosslinking agent to react with said protein and having amolecular weight in a range of about 1000-15,000; and curing saidcomposition on the tissue to bond said composition to the tissue and toprovide a substantive cured matrix.
 262. The method of claim 261 whereinsaid composition is cured to produce the matrix in less than about 10minutes.
 263. The method of claim 261 wherein said composition is curedto produce the matrix in less than about one minute.
 264. The method ofclaim 261 wherein said composition is cured to produce the matrix inabout ten seconds.
 265. The method of claim 261 comprising providing thecomposition to the tissue using a syringe.
 266. The method of claim 261comprising providing the composition to the tissue using a dual syringe.267. The method of claim 261 comprising providing the composition to thetissue using a spray apparatus.
 268. The method of claim 261 wherein thematrix is resorbed.
 269. The method of claim 268 wherein the matrix isresorbed in about four to sixty days.
 270. The method of claim 261comprising curing the composition such that the peel strength of thematrix is about 0.08 lb/in or more.
 271. The method of claim 261comprising curing said composition to provide a cured matrix that has aburst pressure greater than about 10 mmHg.
 272. The method of claim 271wherein the matrix has a burst pressure of about 34 mmHg or greater.273. The method of claim 272 wherein the matrix has a burst pressure ofabout 90 mmHg or greater.
 274. The method of claim 273 wherein thematrix has a burst pressure of about 130 mmHg or greater.
 275. Themethod of claim 261 comprising providing a composition wherein thecrosslinking agent has a molecular weight in a range of about1,000-5,000.
 276. The method of claim 261 comprising providing acomposition wherein the activated leaving group is an N-hydroxy imide.277. The method of claim 276 comprising providing a composition whereinthe activated leaving group is N-hydroxy succinimide.
 278. The method ofclaim 261 further comprising mixing a first mixture and a second mixtureto form the composition and applying said composition to the tissue,wherein the first mixture includes about 20-60 wt/vol % of the proteinin about 0.01-0.25 molar buffer at a pH in a range of about 8.0-11.0 andthe second mixture includes about 50-800 mg/ml of the crosslinking agenthaving a molecular weight in a range of about 1,000-15,000.
 279. Themethod of claim 278 wherein the crosslinking agent is of the formulaG—LM—PEG—LM—G wherein: —PEG— is a diradical fragment represented by theformula—O—(CH ₂ —CH ₂ —O—)_(a) — where a is an integer from 20-300; —LM— is adiradical fragment selected from the group consisting of a carbonatediradical of the formula —C(O)—, a monoester diradical of the formula—(CH ₂)_(b) C(O)— where b is an integer from 1-5, a diester diradical ofthe formula —C(O)—(CH ₂)_(c) —C(O)— where c is an integer from 2-10 andwhere the aliphatic portion of the diradical may be saturated orunsaturated, a dicarbonate diradical of the formula —C(O)—O—(CH ₂)_(d)—O—C(O)— where d is an integer from 2-10, and an oligomeric diradicalrepresented by the formulas —R—C(O)—, —R—C(O)—(CH ₂)_(c) —C(O)—, or—R—C(O)—O—(CH ₂)_(d) —O—(C)O— where c is an integer from 2-10, d is aninteger from 2-10, and R is a polymer or copolymer having 1-10 monomericfragments selected from the group consisting of lactide, glycolide,trimethylene carbonate, caprolactone, and p-dioxanone; and —G is theleaving group selected from the group consisting of succinimidyl,maleimidyl, phthalimidyl, imidazolyl, nitrophenyl, and tresyl.
 280. Themethod of claim 279 wherein the protein in the first mixture is about35-45 wt/vol % serum albumin.
 281. The method of claim 280 wherein thebuffer is 0.05-0.15 molar carbonate/bicarbonate buffer at a pH of about9.0-10.5.
 282. The method of claim 279 wherein the second mixture isabout 50-300 mg/ml of the crosslinking agent having a molecular weightin a range of about 1,000-5,000.
 283. The method of claim 279 whereinthe ratio of a volume of the first mixture to a volume of the secondmixture is in a range of about 1:10 to about 10:1.
 284. The method ofclaim 279 wherein —LM— is an oligomeric diradical —R—C(O)—(CH ₂)_(c)—C(O)— where c is an integer from 2-10 and R is a polymer or copolymerhaving 1-10 monomeric fragments selected from the group consisting oflactide, glycolide, trimethylene carbonate, caprolactone, andp-dioxanone.
 285. The method of claim 279 wherein —G is succinimidyl.286. The method of claim 279 wherein the second mixture includes about300-800 mg/ml of a crosslinking agent having a molecular weight in arange of about 5,000-15,000.
 287. The method of claim 279 wherein —LM—is a diester diradical of the formula —C(O)—(CH ₂)₂ —C(O)—.
 288. Themethod of claim 279 wherein —LM— is a diester diradical of the formula—C(O)—(CH ₂)_(c) —C(O)— where c is an integer from 2-10 and where thealiphatic portion of the diradical may be saturated or unsaturated. 289.The method of claim 279 wherein —LM— is an oligomeric diradical derivedfrom polyglycolic acid.
 290. The method of claim 261 comprising curingthe composition on the tissue to seal the tissue.
 291. The method ofclaim 290 comprising treating tissue to prevent or control a fluid leak.292. The method of claim 291 wherein the fluid leak is a blood leak.293. The method of claim 290 wherein the tissue includes an air leak.294. The method of claim 293 wherein the air leak is in a pulmonarysystem.
 295. The method of claim 261 wherein the composition is providedto tissue at a surgical site.
 296. The method of claims 261 comprisingcuring the composition at the tissue to prevent a tissue adhesion. 297.The method of claim 261 wherein the composition is provided on a surfaceof an internal organ.
 298. The method of claim 261 comprising curing thecomposition to form a matrix to bind tissue.
 299. The method of claim298 wherein the matrix binds tissue together in addition to a suture, astaple, a tape, or a bandage.
 300. The method of claim 261 wherein thecomposition is provided to attach skin grafts.
 301. The method of claim261 wherein the composition is provided to attach adjacent layers oftissue.
 302. The method of claim 261 wherein the composition is providedto position tissue flaps.
 303. The method of claim 261 wherein thecomposition is provided to close gingival flaps.